International Journal of Food and Nutrition Sciences

Vol. 4(2), pp. 065-073, July, 2019. © www.premierpublishers.org. ISSN: 2167-0434

Research Article
Development of Antioxidant Rich Functional Dairy Product
and Study their Total Antioxidant Status Measured by DPPH
Assay
*1Seema Ashraf, Aineeza Fatima2, IfzaMeiraj Khan3, Juveria Aslam4, Zile-E-Noor5, Anum
Liaquat6, S.M. Ghufran Saeed7, Rashida Ali8
1,2,3,4,5,6,8Department
of Food Science and Technology, Jinnah University for Women, Nazimabad, Karachi, 74600,
Pakistan
1,7,8Department of Food Science and Technology, University of Karachi, Karachi, Pakistan

Present investigation has featured the capability of functional dairy product fortified with
essential oil has bearing antiradical properties that decolorize the stable DPPH radical. The
antioxidant activity of Syzygium aromaticum (clove buds) and Cinnamomum verum (cinnamon)
has found to possess inhibitory effect against reactive oxygen species (ROS) due to their high
oxygen radical absorption capacity (ORAC) value. The target of the investigation is to set up the
useful dairy item "coagulated cream", with the fortress of basic oils from S. aromaticum and C.
Verum and blend of both the oils. In addition, determination of the total antioxidant activity of the
product by DPPH (1,1-Diphenyl-2-picrylhydrazyl) assay in three different types of clotted cream
sample prepared along with varying concentrations of S. aromaticum and C. Verum and
combination of both the oils. Additionally, to compare the proximate quality and overall
acceptability of the final product were analysed antioxidant analysis of the product reveals that
the maximum DPPH antioxidant activity percentage was observed in combination of both
essential oils from the S. aromaticum and C. Verum before treatment at 0.25% focus for example
70.9% while the IC50 esteem is 0.029ml at 0.75% focus shows a powerful antioxidant product in
correlation with every single other samples.

Keywords: Syzygium aromaticum, Cinnamomum verum, Antioxidant, functional dairy product, free radicals scavenging,
clotted cream, CLO (clove oil)

INTRODUCTION

In Japan, the term functional food was first coined in 1984.
Functional foods are defined that provide positive
physiological effects with the nutrients. Food and food
products can only be considered functional if together with
the basic nutritional impact it has beneficial effects on the
mankind thus improving the general and physical
conditions and decreasing the risk of development of
diseases (Danik and Emma, 2018). Functional food
contains the proper balance of ingredients which aid us to
function better and effectively in many aspects of our lives,
as well as help us directly in the prevention and treatment
of illness and disease. The benefits of a particular food
component for the human health that people should
struggle to consume a wide variety of food such as to
assure the ingestion of compound such as carotenoids,
flavonoids, fibers, specific fatty acids, minerals,
phytoestrogens, prebiotics and probiotics, soy proteins
and vitamins, among others, in order to reduce the risk of
disease. Pakistan is the one of the top milk producers
according to FAO 2010 in 2007-2008 approximately 42.17
million tons of milk was produced. Majorly about 90-95%
of milk is produced in rural and check -urban regions by
three or four milking animals. Major milk producing animals
are buffalos and cows with 62% and 34% respectively.
*Corresponding Author: Seema Ashraf Ph.D scholar;
Department of Food Science and Technology, University
of Karachi, Karachi, Pakistan.
E-mail: s.seemajuw@gmail.com; Tel: +92-03312819133

Development of Antioxidant Rich Functional Dairy Product and Study their Total Antioxidant Status Measured by DPPH Assay
Ashraf et al. 066
Province of Punjab and Sindh are leading milk producers
with the production of 25.62 million and 9.35 million liters
respectively. Khyber Pakhtoon Khwah produces 4.88
million liters per annum while Baluchistan estimated
produces 0.81 million liters (PDDC, 2006). Clotted cream
can be a value-added product and produce with the
greatest demand in the milk producing districts, Despite
the fact that the product is majorly consumed in European
countries, but it would really contribute in the country’s
exports. Furthermore, it would be an industry returning an
adequate profit to the farmer or the man who undertakes
to pioneer and substantiate the production. It is quite
feasible to prepare the cream on a small scale particularly
on farms, cottage industry or pilot plants. The global
market of functional food promises to have a bright future
according to its compound annual growth rate of 6%
between the periods 2011 to 2015, there is no universally
accepted definition a functional food but according to the
international food information Council functional food or
components of food better provide benefit beyond basic
nutrition. Popular types of product worldwide include
vitamins and daily supplements, functional beverages
such as Sports and energy drinks as well as products
fortified with prebiotics
Clotted cream, also called scalded, clouted Devonshire or
Cornish. It is rich, thick and yellow in color it has cooked or
scalded flavour, created by heating milk which has not
been pasteurized up to the point when a thick creamy layer
appears on its top. Cream contents rises to the surface
during this time and form clots. The milk gets cooled
afterwards and the top layer gets removed. Clotted cream
has grainy or soft crusted surface, with sometimes oily
globules appearance on top. It has a rich cooked milk or
nutty flavour. Although its origin is exciting, the production
of cream is commonly associated with Dairy farm in south
west England and in particular the counties of Cornwall
and devon the current largest commercial producer in the
UK is Roddas at Scorrier, Redruth, Cornwall, which can
produce about 25 tons (25000 kg, 55,000 lb) clotted cream
a day in 1998. Since it has higher fat content so the
consumption of clotted cream on regular basis is thought
to be not suitable for health. Although some researchers
have proved the dairy fat is beneficial in moderate amount.
Syzygium aromaticum commonly called as clove belongs
to the family Myrtaceae. Clove are also known as the
ultimate protective herb and have been used in essential
oil blends such as on Guards and Thieves. Moreira, et
al.,2004 and Ali et al., (2005), reported that the essential
oils from clove possess antimicrobial and anti fungal
activities. Ground cloves has the astounding ORAC value
of 290, 283 units (Kaefer et al., 2008). The strong
antifungal activity of clove oil is due to its high phenolic
content which vaporizers easily (Plaza, et al., 2004). Some
experiments proved that phenols exhibited stronger
antifungal activity compared to other components in
essential oil such as alcohol, aldehydes, ketones and
esters. Eugenol is the main component which is present in
Development of Antioxidant Rich Functional Dairy Product and Study their Total Antioxidant Status Measured by DPPH Assay
S. aromaticum possessed antifungal properties was
reported by (Can et al., 2010) Antioxidant activity of clove
and clove oil are higher than the standard
butylatedhydroxytoluene (BHT). Gallic acid and eugenol
(Kramer,1985) and eugenyl acetate has been identified
from the S. aromaticum and exhibited a antioxidant
activity. Eugenyl acetate and eugenol possessed similar
antioxidant activity with natural antioxidant, alpha-
tocopherol (Vitamin-E),
It is a mild antioxidant when compared with other Essential
oil .It has an ORAC value of 10, 340 micro TE/100 g
Cinnamon essential oils ability to fight cancer (Kaul et al.,
2003) .several studies have reported that the
cinnamanaldehyde from Cinnamomum verum ability to
inhibited the tumor cell proliferation Via Trigger cancer cell
apoptosis (programmed cell death).The widespread
appreciation for healing properties of cinnamon is due to
its medicinal applications.Extracts of cinnamon oil contains
a variety of antimicrobial properties. Studies showed that
the bark extract of Cinnamomum verum were active
against 27 strains of Vibrio cholera and also against
Shigella (Islam et al., 1990). Cinnamon bark oil has
fungicidal and insecticidal properties and also prevents
aflatoxin production.
Clove oil is the most imperative essential oils used for
flavouring every kind of food product, such as sauces,
confectioneries, baked goods, etc. Perfumery and
pharmacy also use clove oil. Particularly clove bud oil used
in pharmaceuticals and perfumery. Also, pickles
confectionery bakery products and sauces. The natural
antioxidants contained in the plant play a vital role in the
treatment of numerous diseases, including diabetes,
atherosclerosis, heart disease and cancer. Secondary
metabolites of plants are essential oils obtained from
flowers, leaves, roots, bark seeds or pericarp. The main
constituent of essential oils is terpenes/Terpenoids
aromatic and aliphatic compounds which are characterize
as a low molecular weight aroma chemical. Generally,
essential oils are comprised of two or three main
component in relatively high concentration (20 to 95%) and
other component present in trace levels that they may be
extracted by steam distillation or hydro distillation
(Majda et al., 2019). At ambient temperature afforded an
essential oil yield of 1.13 %. Upon infrared drying at 45 °C
of Essential oil was lowered by 31%, while convective
drying at 45 °C or 65 °C led to losses of up to 83% as
compared to the essential oil amount that is extracted from
the raw material and dried at ambient temperature.
Following convection drying, essential oil exhibited a loss
of monoterpene hydrocarbons, which was probably due to
chemical rearrangement of compounds or their volatility
with steam. The drying process reduced the content of
monoterpene esters about 13% and increase the amount
of monoterpene alcohols about 4% that this is probably
due to the activation of enzyme belonging to the group of
hydrolysis and hence the change in the quantitative

composition of essential oils. Microwave vacuum drying
lead to Products with a favourable color and an aroma
better than that obtained by conventional methods. The
drying method affects not only the amount of essential oil,
but also often their chemical composition (Nowacka et
al.,2012).
Clotted cream is typically used for flavouring and
garnishing in our food Fortification of clotted cream with
clove and cinnamon essential oil increase the antioxidant
level that will good for cancer patient because eugenol in
clove can protect the liver against disease .it was also
observed that eugenol reserves cellular oxidation which
causes aging and also inflammation. The rationale behind
the present study is to prepare a functional dairy product,
determining antioxidant levels determined by DPPH assay
and comparison of proximate quality and overall
acceptance. DPPH is stable organic nitrogen radical. The
method is simple yet sensitive for determining the
antioxidant compounds in natural products (Ilecholubo et
al., 2017; Nayan et al.,2013). Hydrogen donor is an
antioxidant in the DPPH assay. Radical scavenger
compounds are measured by DPPH.1,1-Diphenyl-2-
picrylhydrazyl converts its color from deep blue-violet to
yellow when scavenged. The scavenging activity of radical
between DPPH and an antioxidant (H-A) can be written as:
MATERIALS AND METHODS
Commercially prepared cream, essential oils of cinnamon
and clove, sucrose, were purchased from the local super
market of Karachi. Distilled water, methanol, 1, 1-
Diphenyl-2-picrylhydrazyl (DPPH), Activated Slica gel
Amyl alcohol, Formalin solution 35%, Methanol, Sodium
hydroxide, sulphuric acid. All chemicals and reagents used
were of analytical grade while food grade oils were used
to conduct the study.
Preparation of clotted cream
The present study was undertaken in the period from July
2017 to January 2018 in the Department of Food Science
& Technology, Jinnah University for women, Karachi,
Pakistan. Commercially prepared cream was used for the
preparation of clotted cream. The preparation of clotted
cream involves the heat treatment up to 88 oC (190oF)
described by (Wilfrid Sadler, 1917) for 8hrs along with
subsequent cooling for 24 hrs. Prior to heat treatment, a
total of 10 samples were prepared. All samples were
prepared on laboratory scale and preserved at 4-5oC.
Preparation and formulation of clotted cream fortified
with essential oils
Three different concentrations; 0.25%, 0.5% and 0.75% of
cream with essential oils of clove, cinnamon and their
combinations were prepared, along with one control
sample. For 0.25%, 250µl clove and Cream with 2gm of
Development of Antioxidant Rich Functional Dairy Product and Study their Total Antioxidant Status Measured by DPPH Assay
Int. J. Food Nutr. Sci. 067
sucrose cinnamon essential oils were added separately in
each container having 50g cream with 2g of sucrose. 0.5%
was prepared by adding 500µl of each essential oil
separately with constant amount of cream and sucrose.
For 0.75%, 750µl clove and cinnamon essential oil were
added. Three different concentrations made with
combinations of clove and cinnamon essential oil. For
combination of 0.25%, 125µl clove +125µl cinnamon
essential oil was added in constant amount of cream and
sucrose. For the combination of 0.5% 250µl clove + 250µl
for cinnamon oil were added in constant amount of cream
and sucrose. Third combination samples i.e. 0.75%, 375µl
clove+375µl cinnamon essential oils were added along
with the constant amount of cream and sucrose.
Chemical analyses
The chemical tests were carried out in triplicates for fat,
protein, carbohydrates, moisture and ash.
Fat content
Fat content was determined by Gerber method described
by Kohler’s measuring method.
Protein content
Protein content was determined by Formol titration method
(AACC, 46-13.01). The protein content was calculated as
follows:
Protein% = (V1-V2) × 1.94
Where:
V1 = burette reading of sample
V2 = burette reading of blank
Moisture content
Moisture content was analyzed by moisture analyzer
AOAC (2017).
Ash content
The ash content was determined according to AOAC
(2017). The ash content was calculated as follows:
Ash% = W1/W0 x 100
Where:
W1 = weight of ash
W0 = weight of sample
Carbohydrate content
The carbohydrate content was determined by subtracting
the summed-up percentage compositions of moisture,
protein, lipid, fibre and ash contents from 100 (Lawal et al.,
2014). It was calculated as follows:
Carbohydrate% = 100 - % moisture - % protein - % lipid -
% ash
Ashraf et al. 068
Antioxidant Capacity Assay
1, 1-Diphenyl-2-picrylhydrazyl (DPPH) free radical
scavenging method was used to assay the free radical
scavenging ability of the fortified samples of clotted cream
(Ilecholubo et al., 2017).
Preparation of sample solution for measurement of
antioxidant level
Briefly, 1g of each prepared samples of clotted cream
(before heat treatment and after heat treatment) was
accurately weighed in centrifuge tubes and homogenized
on vortex with 10 ml of aqueous methanol for about 10 min.
After mixing, the samples were centrifuged at 2000 rpm for
10 minutes and the supernatant was carried out and used
for further analysis.
Experimental procedure for the measurement of free
radical scavenging activity for DPPH
1mM of methanolic DPPH solution was prepared by adding
0.04gm and make up with methanol accurately in 100 ml
volumetric flask. For the DPPH assay, 6 different dilutions
of sample extracts were prepared in the following order i.e.
0 ml, 0.2ml, 0.4ml, 0.6ml, 0.8ml & 1ml with 1ml (300µl) of
DPPH solution. Each dilution then further makes up-to 2ml
by using methanol. The solutions were mixed thoroughly
and were allowed to stand for 30 min in the dark at room
temperature (28±2oC). The absorbance of the resulting
mixture was measured at 517nm using UV-visible
spectrophotometer. The control was prepared by adding
only methanol to DPPH reagent and the analysis followed
as described above. The results were expressed as
percent inhibition (I %) using Equation 3.1
% Inhibition + [A Control – A Test] / [A Control] × 100
Where, A Control: tube containing only methanol; A Test: tube
containing sample extract
Sensory evaluation
Fifteen panellists were selected from among students,
staff and faculty at the Jinnah University for women,
Karachi to evaluate the color, flavour, aroma and overall
acceptability of the functional dairy product in comparison
to the control. They scored the sample on the basis of 9
points hedonic scale, where 9 =like extremely through 5 =
neither like nor dislike to 1 = dislike extremely. The
Panelists were adequately trained to avoid any biasing
during the evaluation of the sample.
Statistical analysis
Analysis of variance of proximate data were done with the
help of Analysis Tool pak program under Microsoft excel
2007. On-way ANOVA for proximate analysis was used to
compare the effects of heat treatments. Means were
compared by Fisher’s Least Significant Test at a
significance level of P> 0.05.
Development of Antioxidant Rich Functional Dairy Product and Study their Total Antioxidant Status Measured by DPPH Assay
DISCUSSION
Proximate analysis of clotted cream prepared with
different concentrations:
Proximate analyses of each prepared samples of clotted
cream were carried out using conventional methods. The
determination of proximate constituents is necessary for
assessing nutritional levels of newly developed functional
product, the analysis revealed important findings.
Data in table 1 shows the proximate composition of clotted
cream prepared with different concentrations of you have
to change and mentioned like S. aromaticum and C. verum
and combination of both oils. The crude fat content ranged
from 40.258 ± 0.114% to 41.823 ± 1.345%. The lowest fat
content was found in concentration 0.75% CLO while the
highest fat content was calculated in concentration 0.5%
CIN sample. (Belgen and Irfan, 2009) analyzed fat content
in 16 kinds of different samples of clotted cream prepared
on a large scale. They analyzed the fat content in samples
ranged from 58.97% - 66.59%. The original milk cannot
have any considerable amount of the percentage of fat in
finished clotted cream. Collectively considerable amount
of fat content was found in clotted cream prepared with
cinnamon oil with all the three concentrations (0.25%,
0.5% and 0.75%), while minimal fat content apparently
was found in clotted cream prepared with clove oil with all
the three concentrations (0.25%, 0.5% and 0.75%). The
significant variations on the basis of sample
concentrations. Since (p> 0.05) therefore, no significant
difference was found on the fat content by changing the
concentration of oils. The protein content ranged from
2.153 ± 0.605% to 3.113 ± 0.220%. The lowest protein
content was found in concentration 0.25% COMB while the
highest protein content was found in concentration 0.5%
CIN sample. The considerable amount of protein content
was found in clotted cream prepared with clove oil with all
the three (0.25%, 0.5% and 0.75%), while minimal protein
content apparently was found in clotted cream prepared
with combination oil with all the three concentrations
(0.25%, 0.5% and 0.75.The carbohydrate content was
found to be in the range from 4.455 ± 0. 627 3% to 5.707
± 0.143%. The lowest carbohydrate content was found in
concentration 0.75% CIN while the highest carbohydrate
content was calculated in concentration 0.5% CLO
sample. Respectively the collectively considerable amount
of ash content was found in clotted cream prepared with
clove and combination oil with all the three concentrations
(0.25%, 0.5% and 0.75%), while minimal carbohydrate
content apparently was found in clotted cream prepared
with cinnamon oil. The moisture content of samples ranges
from 62.362 ± 2.055% to 67.588 ± 1.531%. The lowest
moisture content was found in concentration 0.75% CIN
while the highest moisture content was determined in
concentration 0.25% CLO sample. Moisture content was
found to be higher in all samples with minimum variations
between the samples of all the three concentrations
(0.25%, 0.5% and 0.75%). The ash content was found to

be in the rage from 0.748 ± 0.577% to 1.832 ± 0.262%.
The lowest ash content was found in concentration 0.5%
CLO while the highest ash content was found in
concentration 0.75% CLO sample. Collectively
considerable amount of ash content was found in clotted
cream prepared with combination oil with all the three
concentrations (0.25%, 0.5% and 0.75%), while minimal
ash content apparently was found in clotted cream
prepared with cinnamon oil with all the three
concentrations (0.25%, 0.5% and 0.75%).In contrast the
majority of compositional measures no significant variation
were found between the replicates of clotted cream with
variant concentrations of cinnamon, clove and
combination oils.
Free Radical scavenging activity (DPPH) of clotted
cream
The antioxidant activity by DPPH assay of clotted cream
fortified with essential oils of clove, cinnamon and the
combination of both oils were examined by comparing it
with the control product that contain no essential oil and
the %SCV and IC 50 of product is examined before and
after thermal treatment. Ascorbic acid, which is a natural
antioxidant, is used as standard to compare the
antioxidant potential (Can et al., 2010).
As the standard substance, Ascorbic acid showed
maximum scavenging capacity of 98.790% at 50 μg/ml
concentration and has 6.345 IC50 (Table.2) while clotted
cream fortified with cinnamon oil at a concentration 0.75%
showed maximum scavenging capacity of 68.023% before
thermal treatment while after thermal treatment the
percentage is reduced to 61.279%. The clotted cream
fortified with clove oil at a concentration 0.50% showed
maximum scavenging capacity of 62.209% before thermal
treatment while after thermal treatment the percentage is
reduced to 59.069%. The clotted cream fortified with
combination of both oils at a concentration 0.25% showed
maximum scavenging capacity of 70.930% before thermal
treatment while after thermal treatment the percentage is
reduced to 70.581%. Comparison of antioxidant activity
against DPPH Free radical scavenging shows that sample
having 0.50% cinnamon essential oil is potent antioxidant
before thermal treatment (IC50=0.063 ml decline to
0.339ml ) after thermal treatment as compared to sample
having 0.25 % cinnamon essential oil before thermal
treatment (IC50=0.132 ml decline to 0.346 ml) after thermal
treatment and 0.75%( IC50=0.150 ml to 0.214ml) after
thermal treatment (Table.2). Comparison of antioxidant
activity against DPPH Free radical scavenging shows that
sample having 0.75%clove essential oil is potent
antioxidant before thermal treatment (IC50=0.054 ml
decease into 0.196ml ) after thermal treatment as
compared to sample having 0.50 % clove essential oil
before thermal treatment (IC50=0.191 ml to 0.432 ml)
after thermal treatment and 0.25%( IC50=0.216 ml to
0.403ml) after thermal treatment. The 0.75% combination
of both essential oils is potent antioxidant before thermal
Development of Antioxidant Rich Functional Dairy Product and Study their Total Antioxidant Status Measured by DPPH Assay
Int. J. Food Nutr. Sci. 069
treatment (IC50=0.029 ml decline to 0.369ml) after thermal
treatment as compared to sample having 0.25 %
combination of both essential oils before thermal treatment
(IC50=0.194 ml decline to 0.412 ml) after thermal treatment
and 0.50% (IC50=0.286 ml decline to 0.412ml) after
thermal treatment. Hence the IC50 value of the clotted
cream containing combination of both essential oils at
0.75% concentration before thermal treatment is found to
be best antioxidant because of the lowest IC50 value
i.e.0.029 ml followed by the sample that containing 0.75%
clove oil and cinnamon oil having IC50 0.054ml and
0.063ml respectively. The observed increase of interest in
antioxidant has provided a stimulus for scientists to search
for new drying methods, preserving the highest amount of
biologically active compounds possible, as well as
minimizing changes in the appearance, taste, color, and
texture of the materials (Kaefer and Milner 2008;Milene et
al., 2013).Present study have highlighted the potential of
functional dairy product fortified with essential oil has
bearing antiradical properties that decolourized the stable
DPPH radical. Antioxidant analysis of the product reveals
that the maximum DPPH antioxidant activity percentage
was observed in combination of both essential oils before
heat treatment at 0.25% concentration i.e. 70.9% while the
IC50 value is 0.029ml at 0.75% concentration indicates a
potent antioxidant product in comparison with all other
samples.
Sensory Evaluation of clotted cream
Based on mean results of hedonic scores of clotted
creams prepared with different concentrations of oils
representing in figure 3.3.1, four sensory attributes; color,
flavour, aroma and overall acceptability clotted cream
prepared with different concentrations of oils were
evaluated. The flavour of the cinnamon 0.50% and clove
0.25% were most acceptable. While the most unlike
flavour was recorded for control, followed by cinnamon
0.75%. Cinnamon 0.25%, clove 0.50% and 0.75%,
combination 0.50% and 0.75% was moderately acceptable
essential oils are natural antioxidants, and not only impart
sensory qualities (flavor, aroma) to food, but also improve
its microbiological stability (Wargovich et al., 2001). Most
liked aroma was recorded for cinnamon 0.25% and 0.75%.
While less acceptable aroma was recorded with the
samples of control, clove 0.50%, 0.75% and combination
0.50%. Moderate aroma was found to likeable in
combination 0.25%. Color was not much affected by the oil
concentrations, so no visible variations were found in the
samples. Overall cinnamon 0.50%, clove 0.25% followed
by combination 0.75% was categorized in most preferable
range, while control lies in least acceptable category.
Cinnamon 0.75%, combination 0.25, combination 0.50%
lies in moderate acceptable category showed in table .3.
CONCLUSION
From the above study concludes that the scavenging
activity of all essential oil is greater before thermal
Ashraf et al. 070
treatment and after thermal treatment scavenging % is
reduced, similarly IC50 value of all essential oil is less
before heat treatment and after heat treatment IC50 value
after treatment higher, the maximum scavenging %
observed in clotted cream fortified with combination of both
oils at a concentration 0.25% i.e. 70.930% before thermal
treatment and after thermal treatment maximum
scavenging % observed in clotted cream fortified with
combination of both oils at a concentration 0.25%
i.e70.581%. IC50 value of the clotted cream containing
combination of both essential oils at 0.75% concentration
before thermal treatment shows higher antioxidant
potential as compared to others.
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fistulaLinn. PMCID Volume: 34, Issue Number: 2. Accepted 29 April 2019

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Citation: Ashraf S, Fatima A, Khan I, Aslam J, Zile-E-Noor,
Liaquat A, Ghufran Saeed SM, Ali R. (2019). Development
of Antioxidant Rich Functional Dairy Product and Study
their Total Antioxidant Status Measured by DPPH Assay.
International Journal of Food and Nutrition Sciences. 4(2):
065-073.
29
Copyright: © 2019. Ashraf et al. This is an open-access
article distributed under the terms of the Creative
Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium,
provided the original author and source are cited.

APPENDIX

Figure 1: DPPH free radical scavenging activity of Figure 2: Percentage inhibition of standard at various
clotted cream of control sample concentrations

Table .1: Proximate analysis of clotted cream prepared with different concentrations
Conc. Moisture (%) Ash (%) CHO (%) Crude Fat (%) Crude Protein (%)
Sample A
Control 63.486 ± 1.527 1.105 ± 0.133 6.486± 1.527 40.621 ± 0.266 2.976 ± 0.256
Sample B (CIN)
0.25% 66.020 ± 1.002 1.235 ± 0.353 5.296 ± 0.049 41.731 ± 1.282 2.616 ± 0.662
0.5% 64.617 ± 2.450 1.384 ± 0.08402 5.161 ± 0.883 41.823 ± 1.345 3.113 ± 0.220
0.75% 62.362 ± 2.055 1.079 ± 0.33094 4.455 ± 0.627 41.661 ± 1.314 2.500 ± 0.636
Sample C (CLO)
0.25% 67.588 ± 1.531 1.210 ± 0.292 5.121± 0.711 41.662 ± 1.163 2.937 ± 0.915
0.5% 64.485 ± 2.632 0.748 ± 0.577 5.707± 0.143 41.559 ± 0.271 2.631 ± 0.498
0.75% 64.574 ± 2.284 1.832 ± 0.262 4.693 ± 0.624 40.258 ± 0.114 2.538 ± 0.486
Sample D (COMB)
0.25% 65.989 ± 1.809 1.556 ± 0.113 5.040 ± 0.284 41.343 ± 0.692 2.153 ± 0.605
0.5% 46.561 ± 1.075 1.719 ± 0.334 5.313 ± 0.990 42.075 ± 1.328 2.593 ± 0.147
0.75% 65.632 ± 1.805 1.333 ± 0.127 5.521 ± 0.350 41.699 ± 1.062 2.758 ± 0.244

Development of Antioxidant Rich Functional Dairy Product and Study their Total Antioxidant Status Measured by DPPH Assay
Ashraf et al. 072

Figure 3.1: DPPH free radical scavenging activity of Figure 3.2: DPPH free radical scavenging activity of clotted
clotted cream containing CLO 0.5% cream containing clove oil 0.25%

Figure 3.3: DPPH free radical scavenging activity of clotted cream containing clove oil 0.75%

Figure 4.1: DPPH free radical scavenging activity of Figure 4.2: DPPH free radical scavenging activity of clotted
clotted cream containing cinnamon oil 0.25% cream containing cinnamon oil 0.50%

Development of Antioxidant Rich Functional Dairy Product and Study their Total Antioxidant Status Measured by DPPH Assay
 Int. J. Food Nutr. Sci. 073

Figure 4.3: DPPH free radical scavenging activity of clotted cream containing cinnamon oil 0.75%

Table 2: Data represents values of IC50 of different prepared concerns of Clotted Cream
Std.Conc. IC50 R2
S.no Ascorbic Acid 6.345 0.986
Before After
Sample Conc.
IC 50 IC 50
1. Clove 0.25% 0.215 0.402
2. Clove 0.50% 0.190 0.432
3. Clove 0.75% 0.053 0.195
4. Cinnamon 0.25% 0.132 0.345
5. Cinnamon 0.50% 0.063 0.339
6. Cinnamon 0.75% 0.150 0.213
7. Combination 0.25% 0.193 0.412
8. Combination 0.50% 0.286 0.327
9. Combination 0.75% 0.028 0.368

Table 3: Sensory evaluation of the clotted cream with different concentrations of oils.
Sample Color Flavour Aroma Overall acceptability
Control 7.500 ± 0.707 5.500 ± 0.707 4.000 ± 1.414 5.500 ± 0.707
CIN 0.25% 7.500 ± 0.707 7.500 ± 0.707 8.500 ± 1.414 7.000 ± 1.414
CIN 0.50% 6.500 ± 0.707 8.500 ± 0.707 8.000 ± 1.414 8.000 ± 1.414
CIN 0.75 % 7.000 ± 1.414 5.500 ± 0.707 8.500 ± 1.414 6.000 ± 1.414
CLO 0.25% 6.500 ± 0.707 8.500 ± 0.707 7.500 ± 1.414 8.000 ± 1.414
CLO 0.50% 7.500 ± 0.707 7.500 ± 0.707 6.000 ± 1.414 7.500 ± 0.707
CLO 0.75% 6.500 ± 0.707 8.000 ± 1.414 6.000 ± 1.414 8.000 ± 1.414
COMB 0.25% 7.000 ± 1.414 8.000 ± 1.414 8.000 ± 1.414 6.500 ± 3.536
COMB 0.50% 7.500 ± 0.707 7.000 ± 1.414 5.500 ± 1.414 6.500 ± 2.121
COMB 0.75% 6.500 ± 0.707 7.500 ± 0.707 8.500 ± 1.414 6.000 ± 1.414

Development of Antioxidant Rich Functional Dairy Product and Study their Total Antioxidant Status Measured by DPPH Assay